LOCUS       pSLS.400                5686 bp    DNA     circular     29-SEP-2020
DEFINITION  pJKR-O-mphR ec83RT.
ACCESSION   urn.local...y6-c7qf3rh
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     rep_origin      37..897
                     /dnas_title="p15a"
                     /vntifkey=33
                     /label="p15a"
     primer_bind     complement(44..63)
                     /Mismatches=0
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="1.7"
                     /Tm="58.5"
                     /annotation_group="886345200515000020510a-65a3-4c64-ad8c-1
                     d3919939f6c"
                     /Sequence="TTTCTCCTGCCACATGAAGC"
                     /created_by="primer3"
                     /label="SLS_158"
     terminator      949..993
                     /dnas_title="rrnBT1 terminator"
                     /vntifkey=43
                     /note="rrnB1 transcriptional terminator"
                     /label="rrnBT1 terminator"
     gene            1035..2045
                     /note="spectinomycin resistance gene"
     primer_bind     complement(1039..1058)
                     /Mismatches=0
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="59.0"
                     /annotation_group="88633903519300b961b421-3256-4958-b1f1-6
                     ee2ca2500fb"
                     /Sequence="ATCACCAAGGTAGTCGGCAA"
                     /created_by="primer3"
                     /label="WUR_ORF_rev_SLS_72 "
     primer_bind     1727..1746
                     /Mismatches=0
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="2.2"
                     /Tm="59.4"
                     /annotation_group="88634930903900ce9d114f-8193-48e0-a0f5-f
                     d897c4e1ba2"
                     /Sequence="ACAAATGTACGGCCAGCAAC"
                     /created_by="primer3"
                     /label="aadA_rev_SLS_96 "
     primer_bind     2046..2065
                     /Mismatches=0
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="58.8"
                     /annotation_group="8863309671140001606871-c325-4c80-919c-e
                     7fd67684fa4"
                     /Sequence="GCGAAACGATCCTCATCCTG"
                     /created_by="primer3"
                     /label="SLS_135 "
     terminator      2189..2612
                     /dnas_title="rrnB terminator"
                     /vntifkey=43
                     /label="rrnB terminator"
     primer_bind     complement(2221..2238)
                     /Mismatches=0
                     /%GC="33.3"
                     /Hairpin_Tm="31.5"
                     /Self_Dimer_Tm="None"
                     /Tm="43.9"
                     /annotation_group="886332970382001c799176-66ca-4069-817f-6
                     151603fbb1c"
                     /Sequence="GATTTAATCTGTATCAGG"
                     /created_by="primer3"
                     /label="pBAD Reverse (Genewiz)"
     primer_bind     2550..2570
                     /Mismatches=0
                     /%GC="47.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="6.6"
                     /Tm="58.2"
                     /annotation_group="8863369719530020c1d16d-40cb-4f1e-b7d6-2
                     f55877aa549"
                     /Sequence="GCCAGGCATCAAATTAAGCAG"
                     /created_by="primer3"
                     /label="SLS_105 "
     misc_feature    2612^2613
                     /Original_Bases="TGCTAGCAAGTAAGGCCGAC"
                     /label="TGCTAGCAAGTAAGGCCGAC"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    2613..2664
                     /note="mphR promoter/operator"
     -10_signal      2629..2634
     primer          complement(2637..2666)
                     /created_by="User"
                     /label="pJKR-O-mphR_BB_rev"
     misc_feature    2641
                     /note="transcription initiation site of mph(A)"
     RBS             2667..2692
                     /dnas_title="g10 RBS"
                     /vntifkey=32
                     /label="g10 RBS"
     gene            2693..3631
                     /created_by="User"
                     /label="RT"
     terminator      3654..3695
                     /dnas_title="bidir terminator"
                     /vntifkey=43
                     /note="tetA/orfL bidirectional terminator termination of
                     bla transcript is slightly weaker than opposite direction"
                     /label="bidir terminator"
     primer_bind     3710..3729
                     /Mismatches=0
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="59.0"
                     /annotation_group="88634103919200b723c616-0595-4f76-9eae-4
                     3db44e61554"
                     /Sequence="TACCACAGCTAACACCACGT"
                     /created_by="primer3"
                     /label="SLS_104 "
     promoter        3713..3856
                     /note="proB 12% rrnB P1 Davis et al 2011"
     primer_bind     complement(3806..3825)
                     /Mismatches=0
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="88633492502600f344f832-106c-4723-9bd4-8
                     b244d245408"
                     /Sequence="CCGTTGTGGTCTCCCTGAAT"
                     /created_by="primer3"
                     /label="SLS_103 "
     RBS             3857..3876
                     /note="Strong RBS Hybrid Rinquest SD8 100 and Salis
                     235826"
     misc_feature    3877..4461
                     /note="mphR codon optimized"
     terminator      4470..4575
                     /dnas_title="t0 terminator"
                     /vntifkey=43
                     /label="t0 terminator"
     CDS             complement(4608..5342)
                     /dnas_title="EryR"
                     /vntifkey=4
                     /note="erythromycin resistance from pGK12-nucMCS"
                     /label="EryR"
     primer_bind     5465..5484
                     /modified_by="User"
                     /Mismatches=0
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="17.9"
                     /Tm="58.7"
                     /annotation_group="88634312655300acb8e549-b3bc-4482-9da3-0
                     1f52812e5e5"
                     /Sequence="TGCCACCTGACGTCTAAGAA"
                     /created_by="primer3"
                     /label="VF2"
     terminator      5601..5648
                     /dnas_title="T7 Terminator"
                     /vntifkey=43
                     /label="T7 Terminator"
     primer_bind     complement(5619..5638)
                     /Mismatches=0
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="58.0"
                     /annotation_group="886347231880008802acb2-f91b-4f77-8ee9-f
                     2123400082b"
                     /Sequence="CTCAAGACCCGTTTAGAGGC"
                     /created_by="primer3"
                     /label="SLS_157"
ORIGIN      
        1 tcagatcctt ccgtatttag ccagtatgtt ctctagacgg atcgcttcat gtggcaggag
       61 aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc
      121 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg
      181 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
      241 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
      301 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
      361 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
      421 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
      481 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
      541 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
      601 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
      661 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
      721 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
      781 gaccaaaacg atctcaagaa gatcatctta ttaaggggtc tgacgctcag tggaacgaaa
      841 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccagc
      901 ggtatcatca acaggcttac ccgtcttact gtcaagagga catccggtca aataaaacga
      961 aaggctcagt cgaaagactg ggcctttcgt tttgctgagg agacttaggg acccttcgaa
     1021 ccccagagtc ccgcttattt gccgactacc ttggtgatct cgcctttcac gtagtggaca
     1081 aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc
     1141 tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg
     1201 gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac
     1261 gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag
     1321 gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc
     1381 gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca
     1441 atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca
     1501 aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg
     1561 gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg
     1621 tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa
     1681 tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc
     1741 agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat
     1801 acttcggcga tcaccgcttc cctcatgatg tttaactttg ttttagggcg actgccctgc
     1861 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc
     1921 tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca agccatgaaa
     1981 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
     2041 cgcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag
     2101 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     2161 ggcgccccag ctggcaattc cgacgtctgg ctgttttggc ggatgagaga agattttcag
     2221 cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg
     2281 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
     2341 cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
     2401 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2461 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     2521 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc
     2581 tgacggatgg cctttttgcg tttctacaaa ctggattgaa tataaccgac gtgactgtta
     2641 catttaggtg gctaaacccg tcaacattta actttaagaa ggagatatac atatgagcat
     2701 cgacattgaa acgacacttc aaaaagctta ccccgatttt gacgtacttt taaaatcaag
     2761 accagcgaca cactacaaag tatacaaaat tccaaagcgt acaattggct atcgtatcat
     2821 agctcaacct acgccacgag taaaagcaat acaaagggat ataattgaaa tactcaaaca
     2881 gcatactcat atccacgatg cagccacagc gtatgttgat gggaagaata tattagataa
     2941 cgccaagata catcaaagct cagtttattt attaaagcta gatttggtca atttttttaa
     3001 taaaataaca ccggaacttt tatttaaggc attagctaga caaaaagtag atatttcaga
     3061 taccaacaaa aatttattaa aacaattttg tttttggaat agaacaaaaa gaaaaaatgg
     3121 tgcgttagtt ttaagtgtag gagcaccaag ctctcctttt atatcgaata ttgttatgag
     3181 cagcttcgat gaagagataa gttctttttg caaggaaaat aaaatatctt actcaagata
     3241 tgctgacgac ttaacttttt caacaaatga aagagatgtt ttagggttag ctcatcaaaa
     3301 agtaaaaaca acattaattc gtttttttgg cacgcggatt attattaata ataacaagat
     3361 cgtatattct tctaaagcac ataatcgcca tgtcacagga gtgacgttaa ctaataataa
     3421 taaattatca ctaggaagag aaaggaaaag atatattact tctttagtat ttaaatttaa
     3481 agaaggcaaa ctttcgaatg tagatattaa tcatcttaga ggcttgatcg gttttgctta
     3541 taatattgag ccagctttta ttgaaagact tgagaaaaaa tatggagaat caaccattaa
     3601 gtctatcaag aaatactctg aaggaggata attaattaac ggcactcctc agcaaatata
     3661 atgaccctct tgataaccca agagggcatt ttttaatgcc catggcgttt accacagcta
     3721 acaccacgtc gtccctatct gctgccctag gtctatgagt ggttgctgga taactttacg
     3781 ggcatgcata aggctcgtaa tatatattca gggagaccac aacggtttcc ctctacaaat
     3841 aattttgttt aactttgaaa taaggaggta atacaaatgc cgcgtccgaa actgaaatct
     3901 gacgacgaag ttctggaagc ggcgaccgtt gttctgaaac gttgcggtcc gatcgaattc
     3961 accctgtctg gtgttgcgaa agaagttggt ctgtctcgtg cggcgctgat ccagcgtttc
     4021 accaaccgtg acaccctgct ggttcgtatg atggaacgtg gtgttgaaca ggttcgtcac
     4081 tacctgaacg cgatcccgat cggtgcgggt ccgcagggtc tgtgggaatt cctgcaggtt
     4141 ctggttcgtt ctatgaacac ccgtaacgac ttctctgtta actacctgat ctcttggtac
     4201 gaactgcagg ttccggaact gcgtaccctg gcgatccagc gtaaccgtgc ggttgttgaa
     4261 ggtatccgta aacgtctgcc gccgggtgcg ccggcggcgg cggaactgct gctgcactct
     4321 gttatcgcgg gtgcgaccat gcagtgggcg gttgacccgg acggtgaact ggcggaccac
     4381 gttctggcgc agatcgcggc gatcctgtgc ctgatgttcc cggaacacga cgacttccag
     4441 ctgctgcagg cgcacgcgta ataactagtg cttggattct caccaataaa aaacgcccgg
     4501 cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc
     4561 aacaggagtc caagcgagct cgtaaacttg gtctgacagc ccttaactta cttattaaat
     4621 aatttatagc tattgaaaag agataagaat tgttcaaagc taatattgtt taaatcgtca
     4681 attcctgcat gttttaagga attgttaaat tgattttttg taaatatttt cttgtattct
     4741 ttgttaaccc atttcataac gaaataatta tacttttgtt tatctttgtg tgatattctt
     4801 gatttttttc tacttaatct gataagtgag ctattcactt taggtttagg atgaaaatat
     4861 tctcttggaa ccatacttaa tatagaaata tcaacttctg ccattaaaag taatgccaat
     4921 gagcgttttg tatttaataa tcttttagca aacccgtatt ccacgattaa ataaatctca
     4981 ttagctatac tatcaaaaac aattttgcgt attatatccg tacttatgtt ataaggtata
     5041 ttaccatata ttttatagga ttggttttta ggaaatttaa actgcaatat atccttgttt
     5101 aaaacttgga aattatcgtg atcaacaagt ttattttctg tagttttgca taatttatgg
     5161 tctatttcaa tggcagttac gaaattacac ctctttacta attcaagggt aaaatggcct
     5221 tttcctgagc cgatttcaaa gatattatca tgttcattta atcttatatt tgtcattatt
     5281 ttatctatat tatgttttga agtaataaag ttttgactgt gttttatatt tttctcgttc
     5341 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5401 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5461 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5521 cgtatcacga ggccctttcg tcttcactaa gcgactaaaa aattgaatgt aggaaaccaa
     5581 catgccagtt cgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     5641 gttttttgct gaaacctcag gcatttgaga agcacacggt cacact
//
